LOCUS       sequence-472334-        5619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of bovine arrestin-2 short isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5619)
  AUTHORS   Vishnivetskiy SA, Zhan X, Gurevich VV
  TITLE     Expression of Untagged Arrestins in E. coli and Their Purification.
  JOURNAL   Curr Protoc. 2023 Sep;3(9):e832. doi: 10.1002/cpz1.832.
  PUBMED    37671938
REFERENCE   2  (bases 1 to 5619)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr 
            Protoc."; date: "2023-09"; volume: "3(9):e832. doi"; pages: " 
            10.1002/cpz1.832"
COMMENT     Sequence Label: BtARR2 short
FEATURES             Location/Qualifiers
     source          1..5619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(680..685,686..702,703..709)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    717..733
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     722..744
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    750..819
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             870..896
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     primer_bind     complement(2290..2307)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2460..2546
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2638..2665
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2684..2775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2776..2844,2845..3636)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2994..3013)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3807..4395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4296..4315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4549..4566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4581..4721
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4647..4666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4807..4829
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4907..4984
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             join(4985..5619,1..448)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5004..5023)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
       61 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
      121 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
      181 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
      241 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
      301 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
      361 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      421 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagcgcgaat
      481 tgatctggtt tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc
      541 agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc
      601 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
      661 aatattctga aatgagctgt tgacaattaa tcatccggct cgtataatgt gtggaattgt
      721 gagcggataa caatttcaca caggaaacag cgccgctgag aaaaagcgaa gcggcactgc
      781 tctttaacaa tttatcagac aatctgtgtg ggcactcgac cggaattatc gattaacttt
      841 attattaaaa attaaagagg tatatattaa tgtatcgatt aaataaggag gaataaacca
      901 tgggcgacaa agggacgcgg gtgttcaaga aggcgagccc caatggaaag ctcaccgtct
      961 atctgggaaa gcgggacttt gtggaccaca tcgacctcgt ggagcccgtg gatggagtgg
     1021 ttcttgtgga tccggagtat ctcaaggaga ggagagtcta tgtgacgctg acctgcgcct
     1081 tccgctacgg ccgggaggac ctggatgtcc tgggcctgac ctttcgcaag gacctgtttg
     1141 tggccaacgt gcagtctttc ccgccggccc ctgaggacaa gaagcccctg acgcggctgc
     1201 aggagcgcct catcaagaag ctgggcgagc atgcctaccc tttcaccttt gagatccctc
     1261 cgaacctccc atgctctgtg actttgcagc cgggacctga agatacaggg aaggcctgcg
     1321 gtgtggacta cgaagtgaaa gccttctgtg cggagaacct ggaggagaag atccacaagc
     1381 ggaattctgt gcgcctggtc atccggaagg ttcagtatgc cccagagagg cctggccccc
     1441 agcccacggc cgagaccacc aggcagttcc tcatgtcaga caagcccttg catctggagg
     1501 cctccctgga caaggagatc tactaccacg gagaacccat cagtgtcaac gtccatgtca
     1561 ccaacaacac caacaagacg gtgaagaaga tcaagatctc ggtgcgccag tatgcagaca
     1621 tctgtctgtt caacacagcc cagtacaagt gccctgtggc catggaagag gctgatgaca
     1681 cagtggcacc cagctctacg ttctgcaagg tctacacgct gacccccttc ctggccaaca
     1741 atcgagagaa gcggggcctc gccctggacg ggaagctcaa acacgaggac acgaacctgg
     1801 cctccagcac cctgttgagg gaaggagcca accgggagat cctgggcatc attgtttcct
     1861 acaaagtgaa agtgaagctg gtggtgtctc gtggcggtga tgtggccgtg gaactgcctt
     1921 tcaccctaat gcaccccaag cccaaagagg aacccccaca ccgggaagtt ccagagcacg
     1981 agacgccggt agataccaat ctcatagaac ttgacaccaa cgatgacgac attgtgtttg
     2041 aggactttgc ccgccagaga ctaaaaggca tgaaggatga caaggaggaa gaggaggatg
     2101 gtaccggctc tccgcggctc aacgacagat agactggggc tgccctccct ccgggcagct
     2161 ccaggtccac tctcatgcac taggatgctt gttcgtcttc ttcctgtcct ggctccccct
     2221 cccctttgtt cttccagttt ctaccagggg gaagcttggc tgttttggcg gatgagagaa
     2281 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     2341 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     2401 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     2461 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     2521 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     2581 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     2641 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa
     2701 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2761 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2821 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2881 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2941 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3001 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     3061 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3121 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3181 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3241 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3301 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3361 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3421 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3481 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3541 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3601 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3661 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3721 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3781 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3841 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3901 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3961 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4021 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4081 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4141 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4201 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4261 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4321 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4381 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4441 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4501 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4561 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4621 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4681 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     4741 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4801 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     4861 agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt tacgttgaca ccatcgaatg
     4921 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     4981 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     5041 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     5101 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5161 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     5221 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     5281 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     5341 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     5401 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     5461 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     5521 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     5581 tggctggcat aaatatctca ctcgcaatca aattcagcc
//
